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ImmunologicLaboratoryTestsKris5neKra6s,M.D.

•  Agglu5na5onreac5ons•  DAT

•  IAT•  Immunofluorescence

•  ELISA

•  Westernblot

•  Flowcytometry

ImmunologicLabTestsOutline

•  Whatdoesitmeasure?

•  WheredoestheAg-Abinterac5onoccur?

•  HowistheAg-Abcomplexdetected?

ThingstoRememberAboutEachTest

•  Agglu5na5onreac5ons


•  Detec5onofAginthepa5ent’sspecimen

•  Examples:•  tes5ngforan5bodiestoinfec5ousagents•  tes5ngforHemophilusinfluenzaetypeBcapsularan5geninCSF

Agglu5na5onReac5ons:Purpose

nega5ve posi5ve

•  Detec5onofAbonpa5ent’sredcells

•  AlsocalledthedirectCoombsTest

•  Performedinpa5entswithhemoly5canemia

DAT:Purpose

patient red cells + AHG = agglutination


•  IAT


•  Detec5onofan5bodiestoredcellan5gens

•  AlsocalledtheindirectCoombsTest

•  Performedaspartofpre-transfusiontes5ng

IAT:Purpose

patient Ab + AHG = agglutination donor

RBC = Ab-coated donor RBC +

•  Detec5onofaspecifican5genoran5bodyinapa5ent’sspecimen

•  Examples:•  detec5onofan5geninapa5ent’sbiopsyspecimen•  detec5onofan5bodyinapa5ent’sserum

Immunofluorescence:Purpose

Posi5vefluorescentan5nuclearan5bodytest



•  ELISA


•  Detec5onofan5bodiesinpa5entspecimen

•  Examples:•  homepregnancytests•  HIVtests•  testsforsomecoagula5onfactors,cytokines,andautoan5bodies

ELISA:Purpose



•  ELISA

•  Westernblot


•  Detec5onofan5bodiesinpa5entspecimen

•  Mostcommonexample:HIVtest

WesternBlot:Purpose

Enoughbands=pa5entis“posi5ve”



•  ELISA

•  Westernblot



•  Characteriza5onofcellsize,complexity,an5gens

•  Examples:•  diagnosisofleukemiaandlymphoma•  determina5onofCD4/CD8countsinpa5entswithHIV

FlowCytometry:Purpose



•  ELISA

•  Westernblot



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